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Detection method for target gene

A detection method and gene technology, applied in the field of rapid detection of target genes, can solve the problems of high cost, increased cost of detection products, cumbersome production process of colloidal gold chromatography test paper, etc., so as to reduce the professional level, simplify the process of PCR detection, reduce cost effect

Inactive Publication Date: 2018-06-19
宝瑞源生物技术(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the production process of colloidal gold chromatography test paper is cumbersome, the cost is high, and it is difficult to achieve uniform detection results for different batches of test paper
In addition, the production of colloidal gold chromatography test paper requires special workshops, instruments, materials, etc., which increases the cost of testing products

Method used

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  • Detection method for target gene
  • Detection method for target gene

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] In this example, the exogenous gene Cry1Ab (registered in GENBANK search) of transgenic maize was used as the target gene.

[0022] Design marker primers. Synthesis and labeling steps were synthesized by Shanghai Sangon Bioengineering Co., Ltd. Its sequence is:

[0023] Upstream primer: Digoxigenin-TGT GTG ATT TAT CAT CGA

[0024] Downstream primer: Biotin-CGC TCA GTG GAA CGA AAA CTC.

[0025] The above primers are used to prepare the reaction mixture, the components of which are as follows.

[0026] composition

volume

UNGase

0.5ul

Taq DNA polymerase

1ul

5x Taq PCR buffer

10ul

du-dNTP

4ul

primer-F

0.5ul

primer-R

0.5ul

ddH2O

18.5ul

total 35ul

[0027] Template nucleic acid was extracted using a plant tissue nucleic acid extraction kit. Mix 15ul of the template with the above reaction system, and set a blank control at the same time, the blank template can be double dis...

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Abstract

The invention discloses a detection method for a target gene. An upstream primer and a downstream primer are designed on the target DNA, and digoxin and biotin are marked on 5' end respectively. An PCR amplified reaction is carried out by using the primers, after the reaction is finished, the primers are directly adsorbed and added into an antibody protein solution containing avidin and digoxin for interpretation, the result can be observed through naked eyes, other instrument or equipment does not needed, the detection step is reduced, the risk of pollution is reduced, the detection cost is reduced, and the detection method has relatively high wide universality.

Description

technical field [0001] The invention belongs to the field of nucleic acid detection, in particular to a method for rapidly detecting target genes. Background technique [0002] DNA polymerase chain reaction (PCR) is a classic method to amplify DNA fragments, controlled by thermostable DNA polymerase, oligonucleotide primers and temperature cycles. A large number of DNA fragments can be obtained in a short time. Because of its high sensitivity, simple operation and other advantages, it is widely used in medicine. It is the classic method of nucleic acid testing. [0003] At present, the detection of PCR products includes agarose gel electrophoresis, nucleic acid molecular hybridization and so on. The former needs to make agarose gel, and after electrophoresis, put it on a UV transilluminator for analysis. The operation steps are cumbersome and time-consuming; As a result, although a large number of samples can be tested, the requirements for the experiment are high and pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/686
CPCC12Q1/686C12Q2565/113
Inventor 李雨峰陈立柱杨海侠
Owner 宝瑞源生物技术(北京)有限公司
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